The cooperation between TESCAN, a.s., Brno University of Technology and Institute of Molecular Genetics led into the first prototype of a Multimodal holographic microscope (MHM) - a unique instrument using the incoherent holography for the quantitative phase imaging (1). This microscopic methodology is suitable for non-invasive imaging and characterization of cellular mass and growth dynamics. This provides a powerful quantitative tool for label-free biological cell investigation, without using any adverse markers and toxic substances. MHM enables imaging in dispersion solutions, suppresses the coherent noise significantly and therefore provides high imaging sensitivity. The microscope allows quantitative imaging of the phase difference between the reference and object wave, which monitors dry mass distribution within the cells. In addition, it provides the combination of unique holographic properties with other conventional microscopic techniques and maintains the optimal conditions for investigation of living cells during long-term observations (2).
Our study demonstrates the applicability of a multimodal holographic microscope as a suitable tool for quantitative analysis of the cell cycle. First of all, we compared measurements of the same biological samples using the conventional fluorescence technique and the off-axis holographic technique. We obtained similar information about cellular behaviours at different phases during a full lifecycle of HeLa cells from these two different methods. We calculated distribution of intracellular dry mass and its changes in synchronous cells.
References:
(1) Slabý T., Kolman P., Dostál Z., Antoš M., Lošťák M., Chmelík R.: Off-axis setup taking full advantage of incoherent illumination in coherence-controlled holographic microscope, Optics Express, Vol. 21, Issue 12, pp. 14747-14762 (2013).
(2) Kolman P., Chmelík R.: Coherence-controlled holographic microscope, Optics Express, Vol. 18, Issue 21, pp. 21990 - 22003 (2010).
This study was supported by the grant MPO-Tescan (Reg. No. FR-TI4/660).